Simulation-based inference of differentiation trajectories from RNA velocity fields.

We report Cytopath, a method for trajectory inference that takes advantage of transcriptional activity information from the RNA velocity of single cells to perform trajectory inference. Cytopath performs this task by defining a Markov chain model, simulating an ensemble of possible differentiation trajectories, and constructing a consensus trajectory. We show that Cytopath can recapitulate the topological and molecular characteristics of the differentiation process under study.

Visualizing hierarchies in scRNA-seq data using a density tree-biased autoencoder.

Motivation: Single-cell RNA sequencing (scRNA-seq) allows studying the development of cells in unprecedented detail. Given that many cellular differentiation processes are hierarchical, their scRNA-seq data are expected to be approximately tree-shaped in gene expression space. Inference and representation of this tree structure in two dimensions is highly desirable for biological interpretation and exploratory analysis.

Exhausted CD8 T cells exhibit low and strongly inhibited TCR signaling during chronic LCMV infection.

Chronic viral infections are often associated with impaired CD8 T cell function, referred to as exhaustion. Although the molecular and cellular circuits involved in CD8 T cell exhaustion are well defined, with sustained presence of antigen being one important parameter, how much T cell receptor (TCR) signaling is actually ongoing in vivo during established chronic infection is unclear. Here, we characterize the in vivo TCR signaling of virus-specific exhausted CD8 T cells in a mouse model, leveraging TCR signaling reporter mice in combination with transcriptomics.

Landscape of Exhausted Virus-Specific CD8 T Cells in Chronic LCMV Infection.

A hallmark of chronic infections is the presence of exhausted CD8 T cells, characterized by a distinct transcriptional program compared with functional effector or memory cells, co-expression of multiple inhibitory receptors, and impaired effector function, mainly driven by recurrent T cell receptor engagement. In the context of chronic lymphocytic choriomeningitis virus (LCMV) infection in mice, most studies focused on studying splenic virus-specific CD8 T cells. Here, we provide a detailed characterization of exhausted CD8 T cells isolated from six different tissues during established LCMV infection, using single-cell RNA sequencing.

The type IV pilin PilA couples surface attachment and cell-cycle initiation in .

Understanding how bacteria colonize surfaces and regulate cell-cycle progression in response to cellular adhesion is of fundamental importance. Here, we use transposon sequencing in conjunction with fluorescence resonance energy transfer (FRET) microscopy to uncover the molecular mechanism for how surface sensing drives cell-cycle initiation in We identify the type IV pilin protein PilA as the primary signaling input that couples surface contact to cell-cycle initiation via the second messenger cyclic di-GMP (c-di-GMP). Upon retraction of pili filaments, the monomeric pilin reservoir in the inner membrane is sensed by the 17-amino acid transmembrane helix of PilA to activate the PleC-PleD two-component signaling system, increase cellular c-di-GMP levels, and signal the onset of the cell cycle.

Quantitative Selection Analysis of Bacteriophage φCbK Susceptibility in Caulobacter crescentus.

Classical molecular genetics uses stringent selective conditions to identify mutants with distinct phenotypic responses. Mutations giving rise to less pronounced phenotypes are often missed. However, to gain systems-level insights into complex genetic interaction networks requires genome-wide assignment of quantitative phenotypic traits.